RT2 Easy First Strand Kit

For cDNA synthesis and genomic DNA elimination in RNA samples for use with RT2 Profiler PCR Arrays

Rapid and efficient first-strand cDNA synthesis

Complete elimination of genomic DNA

Results with as little as 25 ng total RNA per reaction

cDNA is immediately ready for real-time PCR

The RT² Easy First Strand Kit provides a rapid and convenient procedure for efficient first strand cDNA synthesis and genomic DNA elimination in RNA samples. Its unique format enables easy parallel processing of multiple RNA samples for reverse transcription. The synthesized cDNA is ready to use in real-time PCR expression analysis of multiple genes when used with RT² SYBR Green Mastermixes, RT² qPCR Primer Assays, or other commercially available primer assays.

The RT2 Easy First Strand Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Principle

The RT2 Easy First Strand Kit provides a rapid and convenient procedure for efficient cDNA synthesis from mRNA for use in gene expression assays.

The kit includes a proprietary procedure to effectively eliminate contaminating genomic DNA from RNA samples before reverse transcription. Random hexamers and oligo-dT primers prime reverse transcription in an unbiased manner, and a reverse transcriptase synthesizes cDNA product with optimal yield and length. A built-in external RNA control helps monitor reverse transcription efficiency and test for enzyme inhibitors. The RT2 First Strand Kit is optimized for real-time PCR-based gene expression analysis using RT2 Profiler PCR Arrays and RT2 qPCR Primer Assays, and can be successfully used with other commercially available gene expression analysis assays.

Procedure

The proprietary Buffer GE2 and sample are mixed and then incubated on a thermal cycler. After addition of enzyme and additional buffer, the sample is then reverse transcribed using any thermal cycler. The transcribed cDNA is immediately ready for use in downstream applications. We recommend RT² Profiler PCR Arrays or RT² qPCR Primer Assays for further analysis.

The RT² Easy First Strand Kit yields results with as little as 25 ng or as much as 5 µg total RNA per reaction. However, the optimal amount of starting material depends on the relative abundance of the transcripts of interest. Lower abundance transcripts require more RNA; higher abundance transcripts require less RNA. Greater amounts of input RNA yield greater number of positive calls (i.e., gene expression detection) in the linear dynamic range of the method.